The goal of this proposal is to translate a set of Mycobacterium tuberculosis (Mtb) derived urinary markers, measured by our new nanotechnology, from adults to pediatric patients. The markers have proven utility in adult patients for active tuberculosis TB screening, and correlate with disease severity. TB kills 1.8 million people yearly. The mortality rate for untreated active TB (1.8 million people/yr) is 68%, compared to 5% following treatment. Most young children presenting to the hospial with possible TB undergo physical examination, and a chest radiograph. As these tests are not specific for active TB, diagnosis is often delayed or missed in children. Hospital TB sputum smear microscopy, mycobacterial culture, and nucleic acid amplification (GeneXpert), remains suboptimal due the low number of pathogens and the inability of children to produce sputum. Gastric aspirates for children are an invasive procedure with a sensitivity of only 30?50%, compared to ~80% in adults, and can take 4-6 weeks. A urine test for active pediatric TB is urgently needed, but has never been possible. We have created and used hydrogel nanocage affinity bait biomarker harvesting technology to detect, for the first time, very low abundance (picogram/mL) TB pathogen shed antigens lipoarabinomannan (LAM) and ESAT6, in the urine of HIV-negative patients hospitalized for sputum culture-proven active pulmonary TB, achieving a high sensitivity (95%) and specificity (80%), compared to diseased and healthy controls, revealing a significant correlation of the urinary concentration of LAM with disease severity. Based on our prior success we propose to use our technology to validate known TB markers, and validate a novel set of TB derived urine markers that bridge adult and pediatric patients. Relevant clinical questions: do children and adult have the same markers that demonstrate high sensitivity and specificity for tuberculosis detection? Can we demonstrate correlation with disease severity in children? We will also apply our technology a low cost urine collection device optimized for TB testing that can be used in underdeveloped global regions. Under Aim 1, we will translate and validate known Mtb derived urinary markers in adult patients (N=200 cases and 200 controls) to pediatric patients (N=100 cases and controls), and use mass spectrometry (MS) to validate known markers and discover/validate new urinary markers. Clinical and microbiological characterization of banked samples includes sputum culture and microscopy, chest imaging, GeneXpert, and TST test. Under Aim 2 we will integrate our nanotechnology into a completely novel one step ?Origami? Urine collection cup for home or field collection which compresses 60 ml of urinary fluid analytes into a flat envelope for mailing, permitting field or home collection, obviating the need for shipment and refrigeration of urine fluid. The origami cup performance will be validated for ESAT6 (TB) and via mass spectrometry analysis in the set of banked urines of Aim 1. A reliable highly sensitive urine screening test for active TB in children, conducted in the field, can reduce the spread of TB, and save children?s lives.